2000年 · 第113卷第02期

Objective To test the potential role of an insertion/deletion polymorphism of angiotensin converting enzyme (ACE) gene on the renoprotective responsiveness to benazepril in type 2 diabetes mellitus with nephropathy. Methods The insertion/deletion polymorphisms of ACE gene were determined by polymerase chain reaction(PCR). Ninety cases with diabetes nephropathy were classified according to the genotype of the ACE gene into 30 cases withⅡ genotypes, 29 with ID and 31 with DD. Mean arterial blood pressure (MABP), urinary albumin excretion rate (UAER), serum creatinine (Scr) and ACE levels were measured before and after the six-month treatment of benazepril (no differences in drug dose between groups). Results At baseline the three groups with different genotypes were similar with regard to sex, age, duration of diabetes, frequency of retinopathy, body mass index, HbA1c, MABP, UAER, serum creatinine, total cholesterol, triglyceride, high density lipoprotein, low density lipoprotein, insulin dose. The dose of benazepril and the number of patients receiving nifedipine after starting ACEI were also similar in the patients with II, ID, DD genotypes, respectively. The serum levels of ACE in patients with DD genotype were the highest, the intermediate with ID and the lowest with II genotype ( P < 0.05). Initiation of 6-month treatment with benazepril induced a significant drop in MABP, UAER and ACE levels in all three groups (P < 0.05), but the reduction was significantly greater in patients with Ⅱ genotype (UAER57.9%, MABP21.4 mm Hg, ACE74.8%), compared to patients with either ID or DD genotype (P < 0.01, respectivly). A mild increase in the serum creatinine was seen in all groups after the initiation of benazepril, but the rate of increase in DD group was the highest (8.9%) and in II group lowest (5.4%) (P < 0.05). A multiple linear regression analysis revealed that the ACE gene polymorphism influenced the decline in albuminuria after initiation of ACE inhibition (R2 = 0.69, P < 0.001). Conclusion The patients with II genotype are particularly susceptible to commonly advocated renoprotective treatment.

Objective To screen for mutation hotspot of Wilson disease (WD) gene in Chinese population. Methods Cycle sequencing was used to detect mutation in exon 8 of WD gene in 30 patients with Wilson disease. Results The same missense mutation, Arg779Leu, was identified in 14 WD patients, four of whom were homozygous and the other heterozygous for this mutation. The frequency of this mutation in Chinese patients was 30%. Conclusion The codon 779 (CCG→CTG) of exon 8 of WD gene was one of mutation hotspots in Chinese. Chin Med J 2000; 113(2): 172-174

Objectives To examine the expression of p16 gene and the clinicopathological characteristics of 31 pituitary adenomas. To investigate the relationship between p16 expression and cell proliferative activity by analyzing the expression of proliferating cell nuclear antigens (PCNA). Methods Immunohistochemistry and in situ hybridization were performed on freshly-prepared paraffin-embedded specimens of 31 pituitary adenomas. Results Varying degrees of overexpression of the p16 gene was found in the tumors specimens. A higher proportion of p16 positive tumor cells was associated with bigger size, more frequent invasion and higher recurrence of the tumors. The expression of p16 was found to be significantly associated with the degree of PCNA expression. Conclusion Though inactivation of the p16 gene is known in wide range of tumors, our study suggested that p16 overexpression might play a role in the development and progression of pituitary adenomas. Chin Med J 2000; 113(2): 162-166

Objective To assess the prevalence of Chlamydia pneumomia DNA in patients with otolaryngic disease. Methods PCR assay was used to detect Chlamydia pneumonia specific Pst I 474 fragment DNA in swabs from patients with acute or subacute pharyngolaryngitis or rhinitis and sinusitis. C. pneumonia specific antibodies in sera were also assayed with microimmuno-fluoresence (MIF). Results About 28% (49/175) of the patients were PCR positive and 25.7% (45/175 ) were MIF antibodies positive. The accordance rate of the two methods was 91.8%. Conclusion It is suggested that the C. pneumonia infection was common in this group of patients and the C. pneumonia Pst I 474 specific PCR was sensitive and specific for detecting C. pneumonia in pharyngolaryngitis or rhinitis and sinusitis. Chin Med J 2000; 113(2): 181-183

Objective To compare the curative effects of three different antifungal regimens in the treatment of cryptococcal meningitis. Methods Twenty-two patients were divided into 3 groups: Group I was given intravenous amphotericin B alone or combination with flucytosine therapy. GroupⅡ received intravenous fluconazole alone or combination with flucytosine. The treatment of GroupⅢ was divided into two steps, where the patients received intrathecal amphotericin B plus intravenous amphotericin B with or without intravenous fluconazole until the mycological culture of cerebrospinal fluid (CSF) turned negative, followed by oral fluconazole or itraconazole as maintenance therapy until direct microscopic examination of CSF showed negative once a week for three consecutive weeks. Results Of the twenty-two patients, 17 (77.3%) were cured, 2 (9.1%) improved, 3 (13.6%) died, and one (4.5%) relapsed. Of the 8 patients in GroupⅠ, 5 were cured, 2 improved, one died and one relapsed; Of the 4 patients in Group Ⅱ, 2 were cured, and 2 died; All the 10 patients in Group Ⅲ were cured without any recurrence. Conclusion The two-step therapeutic regimen may be suited to the treatment of cryptococcal meningitis. Chin Med J 2000; 113(2): 178-180

Objective To investigate the manifestations that heat shock proteins(HSPs) possess in the pathogenesis of asthma and chronic bronchitis. Methods Using reverse transcription-DNA polymerase chain reaction (RT-PCR), we investigated the expression levels of HSP70, HSP90α and HSP90β genes in peripheral blood mononuclear cells(PBMC) at natural state and after heat shock in 14 healthy volunteers, 21 patients with asthma and 18 patients with chronic bronchitis. Results No HSP70 gene but HSP90α and HSP90β expressions were found in non-heat-shocked PBMC of normal control; HSP90α and HSP90β genes may be expressed in PBMC of patients no matter whether they were in acute episode or not. Expression of HSP70 was found in PBMC of patients in acute episodes and of three symptoms-free patients with Aas 3, step 2. No expression of HSP70 gene was found in PBMC of patients in convalescent period but in PBMC of patients in acute episode. HSP90α and HSP90β genes were expressed in PBMC of the two patients groups; After heat shock, expressions of the three genes increased in amount significantly in PBMC of all normal controls and patients. Conclusion Expression of HSP70 gene in PBMC of asthmatic patients and chronic bronchitis was different, indicating that HSPs, especially HSP70 might be involved in the pathogenesis of asthma. Chin Med J 2000; 113(2): 175-177

Objective To determine the antioxidative effects of resveratrol (RES), a polyphenolic compound in red wine, on the oxidation of human low density lipoprotein (LDL) using two different oxidation systems. Methods Oxidation of LDL was induced by adding either Cu2+ or an azo compound. The extent of LDL modification was assessed by measuring the formation of thiobarbituric acid reactive substances (TBARS), the relative electrophoretic mobilities (REM), and the amount of oxidized LDL degradation by macrophages. Results During Cu2+ -induced oxidation, RES reduced TBARS formation in LDL by 70.5%, REM of LDL by 42.3% and the amount of macrophage degradation by 65.7%, respectively. The lag phase of LDL oxidation was also delayed by adding RES both in the copper ion and azo compound-induced oxidation systems. Conclusion RES can protect LDL against both Cu2+ -induced and azo compound-initiated oxidative modification in vitro, which might be due to its free radical scavenging capacity. Chin Med J 2000; 113(2): 99-102

Objective To investigate the expression of mRNA of α1 adrenoceptor in hepatic tissues of cirrhotic patients with portal hypertension. Methods Semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR) were used in 12 hepatic virus B-related cirrhotic patients with portal hypertension and 15 controls from routine liver cutting biopsy for gallstone patients undergoing cholecystectomy who have no history of hypertension. Special α1 adrenoceptor subtype product and internal standard GAPDH product were amplified in the same RT-PCR system simultaneously. The ratio of their integral optical density (IOD) was calculated to stand for the relative expression of the α1 adrenoceptor subtypes. Results The relative mRNA expression of α1a adrenoceptor subtype in hepatic tissues of cirrhotic patients (0.26 ± 0.12) was significantly lower than that of controls (0.86 ±0.38, P < 0.01). The relative mRNA expression of α1b adrenoceptor subtype in hepatic tissues of cirrhotic patients (0.03 ± 0.01) was significantly lower than that of controls (0.23 ± 0.10, P < 0.01). There was no expression of α1d adrenoceptor subtype in hepatic tissues of either cirrhotic patients or controls. The relative mRNA expression of α1a adrenoceptor subtype is significantly higher than that of α1b adrenoceptor subtype in hepatic tissues of both cirrhotic patients and controls (P < 0.01). Conclusion αla adrenoceptor mRNA expression predominated in both normal and cirrhotic hepatic tissues, came in second the expression of α1b adrenoceptor and no expression of α1d adrenoceptor. The decreased expression of mRNA of α1a and α1b adrenoceptor may underlie the decrease of α1 adrenoceptor proteins often seen in the hepatic tissues of cirrhotic patients, and may play an important role in the pathogenesis and maintenance of portal hypertension.

Objective To investigate the possible bone changes in female patients with systemic lupus erythematosus (SLE) induced by long-term administration of Tripterygium Wilfordii Hook. F (TW). Methods 70 female SLE patients were divided into 4 groups accordiog to their drug history: SLE disease control group, corticosteroids treatment group, TW treatment group, and both corticosteroids and TW treatment group. Bone mineral density (BMD) of the lumbar spine 2-4 and biochemical markers of bone turnover were studied. Results Long-term administration of TW could significantly decrease BMD levels in female SLE patients ( P < 0.05). The patients receiving TW for more than 5 years had significantly lower BMD levels compared with those for less than 5 years. The degree of decreased BMD induced by TW was less severe compared with that of prednisone. No significant differences were observed in the biochemical markers of bone turnover among four groups (P > 0.05). Conclusion Long-term administration of TW could decrease BMD levels in women. Osteoporosis may be an important problem for SLE patients treated with TW. Chin Med J 2000; 113(2): 159-161

Objective To investigate the role of human parvovirus B19 (HPV B19) infection in the occurrence of aplastic anemia (AA) and idiopathic thrombocytopenic purpura (ITP). Methods From April 1995 to October 1996 sixty-four patients (40 cases of AA and 24 cases of ITP) at Xijing Hospital were enrolled in this study. Among them, 38 were boys, 26 girls, with age ranging from 1 to 12 years. Control group consisted of 30 healthy children during the same period. From each of them, 2 ml of peripheral blood was taken. For the nested polymerase chain reaction (PCR) study, 50 μl of serum was added to a 100 μl of solution containing 10 μg of proteinase K, which was incubated at 65 °C for 1 hour and then at 95℃ for 10 minutes to inactivate the proteinase K. The formed precipitate was then centrifuged at 14 000 r/m for 15 minutes, and 5 μl of the supernatant was used as the PCR template. The nested PCR amplification was done as follows: 5 μl of each template was added to reach 50 μl of PCR reaction volume, containing 31 μl of bidistilled water, 5 μl of buffer, 200 μmol/L of each dNTP, 1 unit Taq DNA polymerase and 300 ng of each primer. Thirty-five cycles of both first-and second-round PCR amplification were performed under the following conditions: 95℃ for 1 minute, 55℃ for 1.5 minutes, and 72℃ for 1 minute. The amplified fragments were 1112 bp and 104 bp respectively. The second PCR product was electrophoresed on 2% agarose gel, and visualized in UV light. The band position was compared with that of HPV B19-DNA positive control. Results The samples of sera obtained from 40 AA patients, 24 ITP patients and 30 healthy children were analyzed for the HPV B19-DNA. The positive rates of HPV B19-DNA were 25.0 (10/40) and 29.2 (7/24) in AA and ITP patients respectively, which were significantly higher than those in the control group (3.3%, P < 0.05). Conclusion HPV B19 infection is closely related to the onset of AA, as well as ITP, and may be an important cause of childhood AA and ITP.

Objective To determine the asymmetry of the human brain functional activation. Methods With the help of GE Signa Horizon MRI cystem, 14 cases of right-handed volunteers were examined and the blood oxygenation level dependent method was used. The T1-weighted images were obtained with spin echo pulse sequence and the functional imaging (T2*-weighted) was performed using a single shot echo planar imaging pulse sequence. Data analysis was done with Sun Sparc Workstation and by the method of student t test or correlation analysis. Results Most of activation areas were in the left hemisphere under language stimulation, while they were in the right side under music stimulation. Besides, a few brain areas in the contralateral cerebral cortex were also activated under both stimulations. Conclusion The present study supported the hypothesis of the asymmetry of brain functional activation and many brain areas of the cerebral cortex as well as both hemispheres worked in coordination. In addition, it also proved that fMRI is a feasible method in the study of human brain in vivo. Chin Med J 2000; 113(2): 154-158

Objective To develop a new chemical method to produce a monoclonal antibody (MoAb) 520C9/recombinant human interleukin 2 (rhIL-2) conjugate. Methods MoAb 520C9 reactive with the protooncogene c-erbB-2 product P185 was chemically conjugated with rhIL-2 by using a simple two-step method. First, the rhIL-2 was activated by Sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate, a heterobifunctional linker, and N-succinimidyl s-acetylthioacetate was introduced onto 520C9. Then SATA on the 520C9 was reacted with the maleimide group on the activated rhIL-2 to generate 520C9-rhIL-2 immunoconjugate. Results The immunoconjugate retained the antigen binding activity compared to the respective native antibody as determined by an indirect live cell binding assay. The immunoconjugate also possessed IL-2 activity as measured by the standard CTLL-2 cells proliferation assay and the stimulation of human peripheral blood mononuclear cells (PBMCs) into lymphokine-activated killer cells. Conclusion Our method of conjugation of rhIL-2 to 520C9 preserves the binding activity of the antibody and the cytokine function of IL-2. This simple and efficient method of conjugation should be applicable to other types of MoAbs and recombinant cytokines. Chin Med J 2000; 113(2): 151-153

Objective To improve the prognosis and therapeutic efficacy of Waldeyer's ring non-Hodgkin lymphoma (NHL-WR), combined radio-chemotherapy was used to treat the patients with NHL-WR and prognostic factors were analyzed. Methods Ninety patients with stage I ~ IV NHL-WR were treated with combined radio-chemotherapy. 4000 cGy to 6000 cGy were given to Waldeyer's ring structure and involved cervical nodes. Uninvolved low cervical nodes received 3000 cGy to 4000 cGy. The combination chemotherapy consisted of COPP (cyclophosphamide [CTX], vincristine [VCR], procarbazine [PCZ], prednisone [PDN]) or CHOP (CTX, ADM, VCR and PDN). Univariate analysis was performed to determine the prognostic unfavorable factors. Results Five-year overall survival rate was 69.7%for the whole group, and 83.2% for patients with stageⅠ, Ⅱ. In univariate analyses, over survival (OS) rates of the patients with low and intermediate grade (76.1%), with stage Ⅰ, Ⅱ (83.2%), without fever (75.2%) and with performance status (PS) 0, 1 (85.7%) were significantly better than those of the patients with high grade (53.0%), with stage Ⅲ, Ⅳ(24.7%) with fever (38.5%), with PS2 (41.7%), PS 3, 4 (0.0%), respectively (P < 0.05). Conclusions Combined radio-chemotherapy can improve the 5-year survival rate for the patients with NHL-WR, especially for stage Ⅰ, Ⅱ patients. PS≥2, stage Ⅲ, Ⅳ, fever and high grade histology were associated with unfavorable prognosis. Chin Med J 2000; 113(2): 148-150

Objective To reconstruct the human optic canal and its inner structures, and to provide detailed knowledge of this region for optic nerve decompression for further understanding on the pathologic mechanisms of indirect optic nerve injury. Methods Six optic canals and their inner structures were reconstructed using a computer-aided 3-dimensional reconstruction system. Quantitative measurement of the canal wall thickness, bony canal transverse area, optic nerve transverse area, dural sheath transverse area, subarachnoid space transverse area, and subarachnoid space volume were done by means of the computer morphometric analysis system. The detailed spatial relationship among intracanalicular structures were also carefully identified on the 3-D models. Results The thinnest portion of the canal was the middle part of the medial wall (0.45 ± 0.35 mm) and the narrowest space was in the middle part of the optic canal (the transverse area was 18.21 ± 2.50 mm2). The volume of subarachnoid space which can be considered the compensatory space for distention incurred by the hemorrhage, optic nerve edema, or hematoma was 21.16 ± 4.31 mm3. At the cranial opening , the middle part and orbital opening, its transverse area was 4.45 ± 1.12 mm2, 2.68 ± 1.32 mm2 and 1.23 ± 0.83 mm2, respectively. Conclusions Since the compensatory space was limited, even a tiny amount of blood or swelling of the nerve may cause optic nerve compression. Because the narrowest space was in the middle part of the optic canal and the compensatory space for distention gradually decreases from cranial end to orbital end, the middle part and the anterior part of the optic canal and dural sheath are critical in optic nerve decompression. Chin Med J 2000; 113(2): 140-143

Objective To investigate the mechanism of Chinese medicine ligustrazini against thrombosis, and the effects of ligustrazini on plasminogen activator inhibitor (PAI-1) expression in normal endothelial cells and lipopolysaccharide (LPS) stimulated endothelial cells. Methods Human umbilical vein endothelial cells (HUVECs) were cultured by trypsin digestion method. PAI-1 protein in HUVEC conditioned medium was measured by Sandwich enzyme-linked immunosorbent assay (ELISA), and PAI-1 mRNA expression was determined by Northern blot analysis. Using electrophoretic mobility shift assay (EMSA), we observed HUVEC nuclear factor-kappa B (NF-kB) nuclear translocation. Results LPS treatment of cultured HUVECs resulted in a significant increase in PAI-1 protein and mRNA expression by these cells. However, when HUVECs were incubated with LPS plus ligustrazini, the upregulation of PAI-1 by LPS was abated. Moreover, ligustrazini could decrease the basal level of PAI-1 protein and mRNA in HUVECs as compared with control. Nuclear extracts prepared from HUVECs stimulated by LPS demonstrated that binding to the NF-kB oligo nucleotide increased as compared with the unstimulated cells, but ligustrazini did not change those binding in the absence or presence of LPS. Conclusions Ligustrazini inhibited both basal and LPS-induced PAI-1 protein and mRNA expression in endothelial cells, and the modulation of PAI-1 in HUVECs by ligustrazini might have other mechanisms rather than NF-kB pathway. Chin Med J 2000; 113(2): 136-139

Objective To determine the pattern of malalignment and the extent of chondral damage, the patellofemoral joint was evaluated by CT scanning and the findings were proved by arthroscopic examination. Methods In 20 cases, 40 knees were examined by CT scanning with and without quadriceps contraction in order to differentiate between static and dynamic abnormalities. The CT images were analyzed. The arthroscopic examination was carried out for 27 knees in 20 cases. Results Ten knees had static patellar lateral subluxation and 16 knees dynamic lateral subluxation. Eleven knees had static patellar lateral tilt, and 7 knees dynamic tilt. All of the 27 knees showed arthroscopic patellar maltracking and chondral damage. Conclusions The patellofemoral alignment, affection of quadriceps, and the extent of tight retinaculum may be precisely evaluated by CT measurement and proved by arthroscopic examination. Chin Med J 2000 ; 113(2): 133-135

Methods Two effective ribozymes (HCRZNL and HCRZC), targeted to HCV 5' NC region and C region, were designed and synthesized. Eukaryotic expression vectors, pSV2-gpt, CD-SR α, containing either HCRZNC or HCRZC were constructed and transfected into MT-2 cells, Objective To determine the nucleotide sites of hepatitis C virus (HCV), which can be cleaved by ribozymes? and to obtain a highly effective, nontoxic and inexpensive antisense ribozyme specific for HCV. which had been infected by HCV. Quantitative polymerase chain reaction (PCR) and hybridization were used to determine the effect of inhibition of HCV by ribozymes. Results HCRZNC and HCRZC suppressed the replication of HCV by 54.7% and 62.1%, respectively. Furthermore, when both ribozymes were cotransfected into cells, they suppressed replication by 78.8%. Conclusion The specific antisense ribozymes, have strong inhibitory effects on the replication of HCV in cultured cells, and have better effect when used together.